MOLECULAR CLONING AND EXPRESSION ANALYSES OF THE INTERLEUKIN-8 GENE FROM BOSTRYCHUS SINENSIS IN RESPONSE TO IMMUNE STIMULATION
The sustainable development of Bostrychus sinensis aquaculture has been seriously impacted by bacterial infectious diseases.Interleukin-8(IL-8)plays an important role in anti-infective immunity and regulation of inflammatory responses.The cDNA sequence of B.sinensis IL-8(named BsIL-8)was obtained by PCR amplification and clonal sequencing.The cDNA sequence of BsIL-8 is 1 244 bp in length,comprising a 5'-untranslated region(5'-UTR)of 576 bp,an open reading frame(ORF)of 312 bp,and a 3'-UTR of 356 bp.BsIL-8 encodes 103 amino acids,including an 18-amino acid signal peptide and a 62-amino acid SCY domain.The SCY domain contains a CXC motif(Cys30-Arg31-Cys32)and four conserved cysteine residues(Cys30,Cys32,Cys57,and Cys73).Sequence analyses showed that BsIL-8 had the highest sequence similarity and identity with Boleophthalmus pectinirostris IL-8(92.4％and 86.4％,respectively).The RT-PCR results revealed that BsIL-8 was constitutively expressed in the 10 organs/tissues examined in B.sinensis.The head kidney exhibited the highest expression,followed by the liver,while the peripheral blood showed the lowest expression.Following immune stimulation with poly(I∶C),the expression of the BsIL-8 gene was significantly upregulated in the spleen,liver,head kidney,and peripheral blood of B.sinensis.The highest expression levels were observed at 3,12,24,and 24 h post-stimulation,respectively(P＜0.01).After Vibrio parahaemolyticus infection,the expression of the BsIL-8 gene was significantly upregulated in the spleen and peripheral blood of B.sinensis,with the highest expression levels observed at 12 h post-infection(P＜0.01).Conversely,the liver and head kidney exhibited significant downregulation of the gene,with the lowest expression levels detected at 12 h post-infection(P＜0.01).In conclusion,the BsIL-8 gene is involved in the anti-infection immune responses of B.sinensis.
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